Root system responses of hyperaccumulator Solanum nigrum L. to Cd

Purpose

Though phytoremediation is an important technology for remedying heavy metal-contaminated soils, hyperaccumulation mechanism, especially in root, is still less known.

Materials and methods

Pot culture experiment was used to explore the tolerance mechanism of a cadmium (Cd) hyperaccumulator Solanum nigrum L. by determining the main root traits compared to the non-hyperaccumulator Solanum melongena L. (cultivar name Liaoqie 3) in the same plant family.

Results and discussion

The total root lengths, total root surface areas, and total root volumes of S. nigrum were not significantly decreased (p?<?0.05) compared to their controls when Cd spikes were lower than 20 mg kg^–1. However, the abovementioned three factors of S. melongena were significantly decreased (p?<?0.05) when 20 mg kg^–1 of Cd was spiked. By contrast, S. nigrum showed stronger tolerance to Cd. In addition, S. nigrum showed all Cd hyperaccumulator characteristics, i.e., a Cd hyperaccumulator. S. melongena was a non-Cd hyperaccumulator.

Conclusions

These results indicated that root trait can be a factor of hyperaccumulation because of strong tolerance to Cd.     

镉胁迫对龙葵镉的吸收积累及生理响应的影响

为揭示镉(Cd)富集植物龙葵的Cd吸收、积累与光合生理特性间的相关性,采用室内盆栽试验,在不同Cd添加水平下(0,2,5,10,20mg/kg,分别记为Cd0、Cd2、Cd5、Cd10、Cd20),研究了龙葵对重金属Cd的吸收、积累特性及生理响应。结果表明:龙葵对Cd有极强的耐性,在Cd浓度为20.0mg/kg的污染土壤中仍能正常生长;Cd≤10.0mg/kg的污染土壤中龙葵对Cd的富集、转运系数均大于1;但Cd20处理龙葵对Cd的富集和转运能力显著下降。Cd在龙葵中的分布为地上部地下部,并且各处理(Cd0、Cd2、Cd5、Cd10、Cd20)龙葵地上部Cd的积累量也显著高达地下部Cd积累量的10~15倍。同时,随土壤Cd浓度增加不仅引起龙葵对重金属Cd的吸收、转运的变化,也会引起龙葵光合生理指标的相应变化,当土壤Cd含量增加到10mg/kg时,龙葵地上部旗叶光合速率(Pn)、蒸腾速率(Tr)和气孔导度(Gs)最强,显著高于Cd0、Cd2、Cd5处理(p0.05)23.71%~79.80%,当土壤Cd含量持续增加,龙葵的光合强度呈显著下降的趋势。龙葵吸收Cd主要受植物外部因素土壤Cd浓度(SCCD)的影响,与Pn、Tr和Gs等内部因素并无显著相关性。因此,试图通过对龙葵的光合生理调控实现提高龙葵对Cd的吸收、积累并不能达到理想的效果,对揭示龙葵富集Cd的机理及调控机制还需进一步的研究。     

不同光强下龙葵对镉的吸收积累及生理响应

采用土培方法探讨了不同光强（自然光和50%遮光）和镉（Cd）不同添加量（0、 25、 50、 75和100 mg/kg）复合处理下,龙葵对Cd的吸收累积特征,超氧化物歧化酶（SOD）、 过氧化物酶（POD）、 过氧化氢酶（CAT）和抗坏血酸过氧化物酶（APX）活性以及叶绿素、 丙二醛（MDA）、 谷胱甘肽（GSH）和植物螯合肽（PCs）含量等的变化特征。结果表明, 自然光条件下龙葵叶绿素b、 叶绿素a+b以及地上和地下部Cd富集量均显著低于遮光处理; 叶和根的SOD、 POD、 CAT和APX活性在自然光条件下随土壤Cd添加量的增大先升后降,而在遮光处理下则持续升高; 除Cd 25 mg/kg处理时根POD及叶CAT活性在不同光强处理下没有显著差异外,其余Cd添加量处理下SOD、 POD、 CAT和APX活性在自然光条件下均显著高于遮光处理; 自然光条件下龙葵叶和根的MDA含量显著高于遮光处理,而叶和根的GSH含量和叶中PCs含量均显著低于遮光处理。研究结果表明,遮光环境促进了龙葵对Cd的富集,并且减轻了Cd对龙葵的氧化胁迫。     

镉对龙葵幼苗生长和生理指标的影响

采用营养液培养法研究了镉胁迫条件下龙葵幼苗生长、生理响应及镉积累特性。结果表明,镉胁迫下,龙葵幼苗生长受到一定程度的抑制,并且具有浓度效应和时间效应。镉胁迫还导致龙葵叶片色素含量下降。叶绿素a、b和类胡萝卜素平均含量在高浓度镉（150μmol·L^-1）处理条件下分别较对照降低55．5％、63．9％和43．3％。低浓度镉（25μmol·L^-1）处理15d内显著促进龙葵幼苗根系活力,平均根系活力较对照上升10．4％,而高浓度镉处理下,根系活力呈现先升后降的趋势,镉处理10d之后达到峰值;随着镉浓度的升高和胁迫时间的延长,龙葵幼苗叶片相对电导率、丙二醛（MDA）含量和渗透调节物质均呈现显著上升趋势。相对于对照植株,低浓度镉处理下龙葵叶片平均相对电导率、MDA含量、可溶性糖和脯氨酸含量上升17．7％、117．7％、5．6％和95．3％,而高浓度镉处理下上升幅度更大,分别为39．0％、194．6％、56．3％和758．0％。从积累部位来看,镉主要积累在龙葵幼苗地上部,镉含量由高到低依次为叶片〉茎〉根系,高浓度镉胁迫20d之后根茎叶镉含量为5d时的1．73、1．49和1．40倍,分别为1287．25、1718．14和2385．27μg·g^-1DW。     

Mn胁迫对龙葵和小飞蓬生长及叶绿素荧光特性的影响

采用对Mn有明显耐受能力的植物龙葵（Solanum nigrum L.）和小飞蓬（Conyza Canadensis （L.）Cronq）为材料,用水培方法（Mn离子浓度分别为0.005（CK）、2、4、8、16 mmol·L-1）,研究植物生长、叶绿素含量及叶绿素荧光主要参数。结果表明,Mn胁迫下,随着Mn浓度的升高,龙葵和小飞蓬的叶面积、根长、存活率都有不同程度的下降,株高先略高于对照,而后逐渐下降。叶绿素含量随着培养液中Mn含量的增加显著降低。两种植物的最大光化学量子产量（Fv/Fm）、最大荧光（Fm）、同期光合量子产量（Yield）和表观光合电子传递速率（ETR）均随着Mn离子浓度的增加明显降低;两种植物的初始荧光（Fo）均呈先下降再上升趋势,而非光化学荧光淬灭系数（NPQ）有上升趋势,但处理之间NPQ变化差异不明显。试验表明在Mn胁迫下两种植物的光合作用电子传递过程和电子传递速率被抑制。随Mn处理浓度增大,龙葵受Mn胁迫的影响比小飞蓬小,说明其耐Mn水平较小飞蓬高,更适合用于Mn污染地区的植物修复。     

Genetic diversity of the African hexaploid species Solanum scabrum Mill. and Solanum nigrum L. (Solanaceae)

Two hexaploid species of Solanum sect. Solanum are present in Africa: Solanum scabrum and S. nigrum. Solanum scabrum is a widely cultivated species and is used as a leafy vegetable, as a source of medicine and as a source of ink dye. In previous studies a wide range of morphological diversity has been reported in this species and in some studies subspecies have been proposed. Subspecies are also recognized in S. nigrum. However, it has not been established whether or not the morphological differences are reflected at the genomic level. The present study applies AFLPs to study the genetic diversity in S. scabrum and its relationship to geographical provenance, morphological differences and the possible existence of subspecies within S. scabrum and S. nigrum. The data obtained were analyzed with cluster analysis (using UPGMA and NJ). The results indicate that the genetic variation within S. scabrum was higher within accessions than between accessions. Accessions did not cluster according to their geographical provenance, indicating that accessions from different geographical areas were not significantly different genetically. The clustering reflected neither morphological differences nor domestication status (cultivated or wild). The morphological differences exhibited by S. scabrum could be due to selection by farmers for different plant types. The AFLP derived clustering pattern did not segregate the subspecies recognized in S. scabrum and S. nigrum into separate subclusters.     

Application of plant growth-promoting endophytes (PGPE) isolated from Solanum nigrum L. for phytoextraction of Cd-polluted soils

Many plant growth-promoting endophytes (PGPE) can assist their host plants cope with contaminant-induced stress responses, which can improve plant growth. In this study, four heavy metals resistant endophytic bacteria, Serratia nematodiphila LRE07, Enterobacter aerogenes LRE17, Enterobacter sp. LSE04 and Acinetobacter sp. LSE06, were isolated from Cd-hyperaccumulator Solanum nigrum L. grown in metal-polluted soil. Their plant growth promoting properties such as production of 1-aminocyclopropane-1-carboxylic (ACC) deaminase, indole-3-acetic acid (IAA), siderophores and phosphate solubilizing activity were characterized in vitro. When added to the Cd-amended soils, all of these four bacteria significantly increased Cd extraction from the soils. Subsequently, a pot experiment was conducted to elucidate the effects of inoculating of these PGPE on the plant growth and Cd uptake by S. nigrum L. grown in three different levels of Cd-contaminated soils. Results showed that the inoculation with these PGPE not only stimulated the growth of host plant, but also influenced the accumulation of Cd in the root, stem and leaf tissue of S. nigrum L. All four strains could colonize the rhizosphere soil and even some can be found in plant interior tissues. The present observations demonstrated that PGPE were valuable microorganism resource which can be exploited to improve the efficiency of phytoextraction.     

镉对龙葵种子萌发及子叶抗氧化酶活性的影响

龙葵（Solanum nigrumL.）是Cd超累积植物,可用于植物冶金和Cd污染土壤的植物修复。超累积植物的生理基础研究有助于提高其对Cd的富集效率。龙葵种子在含有不同浓度CdCl（20、30、50、100、150、200μmol·L-1和300μmol·L-1）的琼脂培养基上萌发7d。结果表明,龙葵在200μmol·L-1和300μmol·L-1Cd时发芽率显著降低,而低浓度Cd（30～150μmol·L-1）胁迫下无显著差异;在Cd≥30μmol·L-1时,活力指数、发芽势均显著降低,幼苗的生长受到显著抑制。幼苗生长分析表明：在30～150μmol·L-1Cd处理下根长下降约17%～35%,显著低于200～300μmol·L-1Cd处理（下降79%～90%）;株高随Cd浓度的升高逐渐下降。子叶抗氧化酶活性分析表明：100～150μmol·L-1Cd胁迫下,CAT和APX活性显著上升;在Cd胁迫下,SOD活性降低但维持在较高的活性水平。表明龙葵幼苗能忍耐小于150μmol·L-1Cd,抗氧化酶活性在抵抗Cd毒害方面发挥重要作用。     

Effects of chromium stress on the subcellular distribution and chemical form of Ca,Mg, Fe,and Zn in two rice genotypes

A hydroponic experiment was carried out to study effects of chromium (Cr) stress on the subcellular distribution and chemical form of Ca, Mg, Fe, and Zn in two rice genotypes differing in Cr accumulation. The results showed that Ca, Mg, Fe, and Zn ions were mainly located in cell walls and vacuoles in roots. However, large amounts of metal ions were transferred from the vacuole to the nucleus and to other functional organelles in shoots. Chromium concentrations in the nutrient solution of 50 μM and above significantly decreased Ca concentrations in the chloroplast/trophoplast, the nucleus, and in mitochondria. It further increased Mg concentrations in the nucleus and in mitochondria, as well as Zn and Fe concentrations in the chloroplast/trophoplast. These Cr‐induced changes in ion concentrations were associated with a significant reduction in plant biomass. It is suggested that Cr stress interferes with the functions of mineral nutrients in rice plants, thus causing a serious inhibition of plant growth. The chemical forms of the four nutrients were determined by successive extraction. Except for Ca, which was mainly chelated with insoluble phosphate and oxalic acid, Mg, Zn, and Fe were extractable by 80% ethanol, d‐H₂O, and 1μM NaCl. The results indicated that these low–molecular weight compounds, such as organic acids and amino acids, may play an important role in deposition and translocation of Mg, Zn, and Fe in the xylem system of rice plants.     

Taxonomic status and geographic provenance of germplasm accessions in the Solanum nigrum L. complex: AFLP data

Amplified Fragment Length Polymorphisms (AFLPs) were employed within the taxonomically difficult Solanum nigrum L. complex in order to characterize the genetic diversity present in a collection of the Gatersleben Genebank, to classify taxonomically unknown material, and to correlate the clustering of the examined accessions with their geographic origin. The results from AFLP analyses using two primer combinations on 44 entries from five species led to the detection of four major clusters, simultaneously uncovering significant differences in the levels of genetic diversity within or between species. S. americanumexhibited the highest infraspecific variation despite close geographic origins, simultaneously being placed in a clearly separated cluster in comparison to the other examined species of the complex. In addition, these other species showed even less interspecific variation than was found at the infraspecific level in S. americanum. In terms of taxonomy, the application of AFLPs helped in the classification of 13 black nightshade accessions formerly only listed as Solanum sp. This also was confirmed by morphological determination. Furthermore, one accession formerly classified as S. physalifolium var. nitidibaccatum i) clustered with AFLPs and ii) was identified morphologically as S. villosum. Contradictory classifications remain for two further entries from the same species, found within the S. nigrum cluster after AFLP analyses, while belonging to S. physalifolium var. nitidibaccatum according to herbarium specimen. Finally, as indicated by the information on provenance in geographically separated subclusters in S. americanum and partially in S. villosum, clues on the currently unknown origin of accessions from the genebank seem feasible by AFLP data.     

Interaction of Cd-hyperaccumulator Solanum nigrum L. and functional endophyte Pseudomonas sp. Lk9 on soil heavy metals uptake

Bioaugmentation is a promising method for assisting phytoextraction of heavy metals from contaminated soil, and the development of bioaugmentation-assisted phytoextraction requires the understanding of the mechanism involved in the interaction between plants and inocula. In this study, a pot study was conducted to evaluate the effect of bacterial endophyte Pseudomonas sp. Lk9 which can produce biosurfactants, siderophores and organic acids on the growth and metal uptake of Cd-hyperaccumulator Solanum nigrum L. growing in multi-metal-contaminated soil. The results revealed that Lk9 inoculation could improve soil Fe and P mineral nutrition supplies, enhance soil heavy metal availability, and affect host-mediated low-molecular-weight organic acids secretion, thereby significantly increasing S. nigrum shoot dry biomass by 14% and the total of Cd by 46.6%, Zn by 16.4% and Cu by 16.0% accumulated in aerial parts, compared to those of non-inoculated control. The assessment of phytoextraction showed that Lk9 inoculation elevated the bioaccumulation factor of Cd (28.9%) and phytoextraction rates of all metals (17.4%, 48.6% and 104.6% for Cd, Zn and Cu, respectively), while the translocation factors had negligible difference between Lk9 inoculation (3.30, 0.50 and 0.40 for Cd, Zn and Cu, respectively) and non-inoculated control (2.95, 0.53 and 0.42 for Cd, Zn and Cu, respectively). It was also found that the symbiotic association between S. nigrum and Lk9 significantly increased the soil microbial biomass C by 39.2% and acid phosphatase activity by 28.6% compared to those in S. nigrum without Lk9. This study would provide a new insight into the bioaugmentation-assisted phytoextraction of heavy metal-contaminated soils.     

水稻突变体对镉的吸收及其亚细胞分布和化学形态特点

Wild-type （Zhonghua 11） and mutant rice （Oryza sativa L.） plants were used to investigate the effect of cadmium （Cd） application on biomass production, to characterize the influx of Cd from roots to shoots, and to determine the form, content, and subcellular distribution of Cd in the roots, leaf sheaths, and leaves of the rice plants. Seedlings were cultivated in a nutrient solution and were treated with 0.5 mmol L^-1 of Cd^2＋ for 14 d. The sensitivity of rice plants to Cd toxicity was tested by studying the changes in biomass production and by observing the onset of toxicity symptoms in the plants. Both the wild-type and mutant rice plants developed symptoms of Cd stress. In addition, Cd application significantly （P ≤ 0.01） decreased dry matter production of roots, leaf sheaths, and leaves of both types, especially the mutant. The Cd content in roots of the mutant was significantly （P ≤0.05） higher than that of the wild-type rice. However, there was no significant difference in the Cd content of roots, leaf sheaths, and leaves between the wild-type and mutant rice. Most of the Cd was bound to the cell wall of the roots, leaf sheaths, and leaves, and the mutant had greater Cd content in cell organelles than the wild type. The uneven subcellular distribution could be responsible for the Cd sensitivity of the mutant rice. Furthermore, different chemical forms of Cd were found to occur in the roots, leaf sheaths, and leaves of both types of rice plants. Ethanol-, water-, and NaCl-extractable Cd had greater toxicity than the other forms of Cd and induced stunted growth and chlorosis in the plants. The high Cd content of the toxic forms of Cd in the cell organelles could seriously damage the cells and the metabolic processes in mutant rice plants.     

The effects of Fe deficiency on low molecular weight organic acid exudation and cadmium uptake by Solanum nigrum L.

Abstract

The influence of Fe-deficiency on the root exudation of low molecular weight organic acids (LMWOAs), pH alteration and cadmium (Cd) accumulation and translocation were investigated in morel (Solanum nigrum L.) in hydroponic culture experiments. Tartaric, citric, malic and acetic acids were monitored because these acids were abundant and often detected as root exudates. Results showed that Fe-deficient plants excreted large amounts of LMWOAs in comparison with Fe-sufficient plants across all Cd treatments (p <0.05). In both cases the concentrations of the four organic acids were tartaric > citric > malic > acetic. The results showed that the Fe-deficient plants with higher concentrations of LMWOAs accumulated more Cd (p <0.05) and induced a decrease in solution pH compared with the Fe-sufficient plants. Cadmium accumulation in the Fe-deficient and Fe-sufficient plants had significant positive correlations with the exudation of malic and acetic acids (p <0.05 and p<0.01). Cadmium accumulation in the Fe-sufficient plants had a significant (p<0.01) positive correlation with the exudation of tartaric acid, whereas there was a negative correlation (p<0.01) between Cd accumulation and the exudation of tartaric acid in the Fe-deficient plants. No significant correlation between the exudation of citric acid and Cd accumulation was obtained. Our results indicate that Fe-deficiency could induce Cd accumulation and translocation through an increase of LMWOAs exudation and pH alteration, both of which enhance Cd bioavailability.     

Cancer-preventive peptide lunasin from Solanum nigrum L. inhibits acetylation of core histones H3 and H4 and phosphorylation of retinoblastoma protein (Rb)

Lunasin, a unique 43 amino acid, 4.8 kDa cancer-chemopreventive peptide initially reported in soybean and now found in barley and wheat, has been shown to be cancer-chemopreventive in mammalian cells and in a skin cancer mouse model against oncogenes and chemical carcinogens. To identify bioactive components in traditional herbal medicines and in search for new sources of lunasin, we report here the properties of lunasin from Solanum nigrum L. (SNL), a plant indigenous to northeast Asia. Lunasin was screened in the crude extracts of five varieties of the medicinal plants of Solanaceae origin and seven other major herbal plants. An in vitro digestion stability assay for measuring bioavailability was carried out on SNL crude protein and autoclaved SNL using pepsin and pancreatin. A nonradioactive histone acetyltransferase (HAT) assay and HAT activity colorimetric assay were used to measure the inhibition of core histone acetylation. The inhibitory effect of lunasin on the phosphorylation of retinoblastoma protein (Rb) was determined by immunoblotting against phospho-Rb. Lunasin isolated from autoclaved SNL inhibited core histone H3 and H4 acetylation, the activities of the HATs, and the phosphorylation of the Rb protein. Lunasin in the crude protein and in the autoclaved crude protein was very stable to pepsin and pancreatin in vitro digestion, while the synthetic pure lunasin was digested at 2 min after the reaction. We conclude that lunasin is a bioactive and bioavailable component in SNL and that consumption of SNL may play an important role in cancer prevention.     

硫对超积累东南景天镉累积、亚细胞分布和化学形态的影响

采用差速离心技术和化学试剂逐步提取法研究了硫对超积累东南景天镉累积、亚细胞分布和化学形态的影响。结果表明,增硫处理（S2和S3）显著提高超积累东南景天根、茎和叶的镉含量、累积量及整株累积总量。镉在超积累东南景天根、茎和叶中的含量和分配比例为F1（细胞壁）F3（可溶性部分）F2（细胞器与膜）,细胞壁（F1）是Cd在超积累东南景天细胞内的主要结合位点;超积累东南景天根、茎和叶F1、F2、F3组分中的镉含量随着硫水平的增加而增加,但分配比例变化不一致;超积累东南景天植物体内镉形态以氯化钠提取态（FNacl）、醋酸提取态（FHAc）和水提取态（FW）占优势。增施硫处理,提高超积累东南景天根FNacl、FHAc和FW提取态镉含量和分配比例,降低FE和FHCl提取态镉含量和分配比例;茎FNacl和FHAc提取态镉含量和分配比例增加,FW、FE和FHCl提取态镉分配比例降低;叶FNacl、FHAc和FW提取态镉的含量增加,但对其分配比例影响不大。     

Removal of Cu and Cd from water by Bacopa monnieri L.

Copper and Cd accumulation was determined in Bacopa monnieri, a common aquatic plant. Plants showed a capability to accumulate both metals in single and mixed metal treatments. Copper accumulation was stimulated by the presence of Cd whereas uptake of Cd was inhibited by Cu. The plants showed high concentration factors for both metals. The results suggest the possibility of using this plant for mitigating Cu and Cd pollution in the aquatic environment.     

Silicon influences cadmium translocation by altering subcellular distribution and chemical forms of cadmium in peanut roots

Effects of silicon (Si) on subcellular distribution and chemical forms of Cd in two contrasting peanut cultivars, Qishan 208 (low seed Cd cultivar) and Haihua 1 (high seed Cd cultivar), were investigated by a hydroponics experiment at low Cd level (0.2 μM CdCl₂). Two cultivars differ in Cd translocation, subcellular distribution and chemical forms. In comparison to Qishan 208, Haihua 1 shows higher translocation factors (TFs); this may be resulted from higher Cd in the soluble fraction, and a larger proportion of inorganic Cd (extracted by 80% ethanol) and water-soluble Cd (extracted by d-H₂O) in roots. Pretreatment with Si decreased Cd in the cell wall, and enhanced Cd in the soluble fraction for both cultivars. However, effects of Si on chemical forms and TFs are cultivar dependent. Si enhances Cd translocation and the d-H₂O extractable Cd in Haihua 1. Positive correlations were observed between TF and Cd in the soluble fraction (r = 0.71, P < 0.05), and between TF and d-H₂O extractable Cd in root (r = 0.89, P < 0.001). Therefore, larger proportion of d-H₂O extractable Cd may be responsible for high TF in Si pretreated plants of Haihua 1.     

低温胁迫下枇杷叶片细胞超微结构及膜透性和保护酶活性的变化

以盆栽"解放钟"嫁接的枇杷结果小苗(Eriobotrya japonica Lindl.)为试材, 采用人工降温的方法, 研究低温胁迫下枇杷叶片细胞超微结构及膜透性和保护酶活性的变化.结果表明, 7 ℃、2 ℃低温处理时, 枇杷叶片大多数细胞器基本无损, 排列有序, PMP、MDA含量先上升后下降, 保护酶活性先增大后减小, 表明枇杷小苗对7 ℃、2 ℃低温逆境有一定的适应能力;-2 ℃低温胁迫叶绿体双层膜结构破坏严重, 类囊体垛叠程度很低, 形不成典型的基粒, 还可见线粒体双层膜结构, 但己经没有明显的内脊, 内部呈小泡化, PMP、MDA含量呈一直上升的趋势, 保护酶活性受到抑制, 表明枇杷结果小苗已经受到冻害;-7 ℃低温胁迫细胞质膜已经破裂, 原生质体浓缩, 叶绿体扭曲变形、相互融合, 液泡破裂, 线粒体膜结构受损, 脊消失, PMP、MDA含量大幅度增加, 保护酶活性受到严重抑制, 表明枇杷营养生长已经受到极为严重的伤害.由此可见, 枇杷结果小苗在0 ℃以下受冻严重, 营养生长受阻, 从而影响枇杷正常生长以及产量、品质的提高.